甲绿—[口派]啷咛法对于核蛋白分化染色的改变

MODIFICATON OF METHYL GREEN-PYRONIN TECHNIQUE FOR DIFFERENTIAL STAIN OF NUCLEIC ACID

一.历史1901年Pappenhein开始用甲绿(Methyl green)及[口派]啷咛G牌(Pyronin G)二种染料,各以1%酒精或水溶液染核及细胞质。1910年Unna改用酸性溶液(0.5%石碳酸),因为甲绿对碱性极敏感,必须在中性或酸性溶液中才能染上[1]。直至1940年以前,此法只用作染细胞核的染色质成绿、蓝绿或蓝紫色.

The methyl green-pyronin technique has become a popular method in histochemistrysince Brachet’s application for differential stain of desoxyriboncleic and ribonucleic acidsin 1940.Due to variation in concentration of different brands of pyronin and variationin stainability of different tissues,one often has to spend considerable dyes before theirright proportion is obtained.In the experiment under discussion the results were unsatis-factory when stained in the stock solutions of methyl green(1%E.Merk,in distilledwater containing 0.25%phenol or in 0.1 M sodium acetate buffer,pH 4.1)and pyronin(1%,National Aniline Dye Co.,in the same medium as for methyl green)either separatelyor in a mixture in various proportions.On the other hand,it was found to be much easierto handle in a diluted mixture.To each 40 cc.of distilled water or 0.1 M acetate buffersolution,pH 4.1,was added 20-25 drops of the methyl green and pyronin stock solutionsrespectively,stain overnight,blot and destain in pure acetone for 15-30 seconds.The timeshould be shortened when sections were thinner than 10μ.The destaining could bemuch faster when the acetone absorbed atmosphoric moisture.To overcome the difficulty of obtaining complete dehydration,it was found thatimmersing sections either in dioxan or castor oil before in xylol gave good results.Therate of destaining in acetont could be lowered by adding 1/3 of either dioxan or castoroil into it.